Pharmaceutical composition containing silybin and ve

ABSTRACT

A pharmaceutical composition is prepared from the following raw materials (in parts by weight): silybin (8.75-60), phospholipid (15-65), a Pu&#39;er tea extract (25-200), and vitamin E (6.25-40). The composition has the effect of treating non-alcoholic fatty liver diseases.

TECHNICAL FIELD

The present invention relates to the technical field of medicines, andparticularly to a pharmaceutical composition containing silybin for thetreatment of hepatopathy.

BACKGROUND ART

In the late 1960s and 1980s, the pharmaceutists of West Germany with H.wagner as representative extracted the active ingredient from the fruitof the Silybummarianum, which is named as silymarin, a new class offlavonoid having a C-9 substituents, i.e., a flavonoid lignans condensedwith a dihydroflavonol and a phenylpropanoid derivative.Silybin(silybinin) is one of the main components of silymarin.Pharmacological and toxicological studies have shown that silybin hasthe effects of protecting and stabilizing the hepatocyte membrane,promoting the recovery of hepatocyte and improving the liver function.Silybin has different levels of protection and treatment effects onvarious types of hepatic injury caused by hepatic poisons such as carbontetrachloride, thioacetamide, hydrosycholine, phalloidine, mucronatine,etc. And silybin can be used for treating acute and chronic hepatitis,early hepatocirrhosis, fatty liver, toxic or drug-induced hepatopathy.

The silybin is poor in water solubility and common organic solvents,resulting in low bioavailability and thereby affecting the clinicalefficacy. To improve the bioavilability thereof, domestic and externalpharmacy workers have made substantial amounts of work. The measures toimprove the absorption of poorly soluble drugs axe typically superfinegrinding, salinization, and the addition of cosolvent, etc. In recentyears, the studies have shown that the dissolution and bioavailabilityare greatly improved by the methods of formulating into cyclodextrininclusion compound, solid dispersion, synthetic phospholipid complex andformulating into different dosage forms.

From the perspective of solid preparation, the phospholipid complex is amore specific solid dispersion, which has a fixed melting point, is amolecular compound (complex) whose chemical nature is more stable anddifferent from the compound of drug and phospholipid, such compoundsvaries with the types of phospholipid and ratios of drug tophospholipid, and a phospholipid molecule can be bound with a differentnumber of drug molecules. Deduced from the spectroscopy characteristicsof the complex, the drug has a strong interaction with the polar groupsof the phospholipid, which inhibits the free rotation of the singlechains in the molecule, whereas the two long fatty acid chains of thephospholipid do not participate in the complex reaction and are free toshift and wrap the polar portions of the phospholipid to form alipophilic surface, so that the complex shows strong lipid solubility.The complex changes the physiochemical properties of drug, and thusincreases the lipid solubility of the drugs and reduces the walersolubility of the drags, and promotes the combination of drug moleculesand cell membranes to improve the absorption and increases thebioavailability of the drug.

Pu'er tea is a unique and famous tea in Yunnan province. The localityhas moderate climate, abundant rainfall and is mist-shrouded, Pu'er teais divided into two series by Yunan big leaf species sun-dry tea andreprocessing thereof: the enzymic Pu'er tea by directly re-processinginto the finished product and the enzymic Pu'er tea by re-processingafter the artificial accelerated fermentation, and the patterns of whichare divided into loose tea and compressed tea; natural aging process isalso persistently carried out after the finished products, with theunique qualities gets better.

Pu'er tea is the only post-fermented tea, and substances harmful to thehuman body such as theophylline, tea polyphenols are degaded in the longprocess of fermentation, so the product is mild, does not stimulate thebody, and also can promote metabolism, accelerate the digestion andtransformation of fats and toxins in the body. For the problems ofobesity and three-hypes which are puzzling urbanites, Pu'er tea can playa good mitigation, effect, such as expelling of toxin, nourishing thestomach, anti-inflammatory, reducing the cholesterol, off lipid andremoving grease, cosmetic slimming. Modern technologies show feat Pu'ertea can improve insulin resistance, regulate levels of blood lipid andleptin, etc., and can block the fat accumulation of hepatic parenchymalcell caused by insulin, resistance to some extent.

Non-alcoholic fatty liver disease (NAFLD) is a metabolic stress-inducedhepatic injury that is closely related to insulin resistance andgenetic, susceptibility, the pathological changes of which are similarto alcoholic fatty liver disease. NAFLD is a clinicopathologicalsyndrome characterized by steatosis and fat storage of hepatocytes inthe hepatic lobule but without history of alcohol abuse. NAFLD showsdifferent degrees of hepatic lesion, from simple fatty liver without anyinflammation to severe inflammatory response of severe fibrosis and evencirrhosis, mainly includes 3 types: simple fatty liver, steatohepatitis,fatty cirrhosis.

Non-alcoholic, fatty liver disease treatment:

-   1. Prevention of protopathies or associated risk factors.-   2. Basal treatment: developing a reasonable energy intake and diet    adjustment, taking moderate aerobic exercises, correcting bad.    lifestyles and behaviors.-   3. Avoiding aggravating hepatic injury: preventing a sharp decline    in weight, drug abuse and other factors that may induce exacerbation    of hepatopathy.-   4. Weight loss: requiring all NAFLD patients who are overweight, and    have visceral obesity and rapid weight gain in the short term to    change the lifestyles to control weight and reduce waist    circumference. Basal treatment for 6 months, weight loss <0.45 kg    per month, or body mass index (BMI)>27 kg/m² combined with blood    lipid, blood glucose, blood pressure and other indicators of more    than two abnormalities may consider adding sibutramine or orlistat    and other obesity drugs, weight loss per week should not exceed 1.2    Kg (children do not exceed 0.5 Kg per week); BMI>40 kg/m² or BMI>35    kg/m² combined wife sleep apnea syndrome and other obesity-related    diseases, may consider the proximal end gastric bypass procedures to    lose weight (II-1, II-2, II-3, III).-   5. Insulin sensitizer: combined with type 2 diabetes, impaired    glucose tolerance, fasting plasma glucose and visceral obesity, may    consider the application of metformin and thiazolidinediones in    order to improve insulin resistance and control of blood glucose    (II-1, II-2, II-3).-   6. Hypolipidemic agents: dyslipidemia, with basal treatment and (or)    application of weight loss and hypoglycemic drags for more than 3-6    months, is still mixed with hyperlipidemia or hyperlipidemia,    combined with more than 2 risk factors, should consider adding the    use of fibrates, statins or probucol and other hypolipidemic drugs    (II-1, II-2, II-3).-   7. Drugs for hepatopathy: NAFLD associated with hepatic dysfunction,    metabolic syndrome, 3-6 months after basal treatment remains    ineffective, and liver biopsy shows NASH and chronic progression of    the course of the disease, the drug auxiliary treatment for    hepatopathy can be used with antioxidant, anti-inflammatory,    anti-fibrosis, and related drugs (II-1, II-2, II-3, III) such as    polyene phosphatidylcholine, vitamin E, silymarin and    ursodeoxycholic acid can be rationally chosen according to drug    performance, disease activity and stage of fee disease, but    multi-drugs should not be applied simultaneously.-   8. Liver transplantation: mainly for NASH-related end-stage    hepatopathy and some cryptogenic J hepatocirrhosis, and fee    metabolic condition (III) should be screened before liver    transplantation. BMI>40 kg/m² is contraindication to liver    transplantation (III).

The above treatments have not been used by being mixed together, such asa combination of hypoglycemic and hepatopathy drugs, or a combination oflipid-lowering and hepatopathy drugs. Therefore, the search for apharmaceutical with a variety of health-promoting functions cannot wait,

SUMMARY OF THE INVENTION

In order to solve fee above technical problems, the present inventionprovides a pharmaceutical composition and a preparation thereof which istherapeutically effective for non-alcoholic fatty-liver diseases, andalso has a synergistic effect on the treatment of non-alcoholic fattyliver diseases when the four are combined in use.

The present invention provides the methods for preparing thepharmaceutical composition and the preparation thereof.

The present invention is achieved by the fallowing technical solutions:

A pharmaceutical composition is prepared from the following bulk drugsby weight ratio:

-   -   8.75-60 parts of silybin,    -   15-65 parts of phospholipid,    -   25-150 parts of Pu'er tea extract, and    -   6.25-40 parts of vitamin E.

It is preferably prepared from the following bulk drugs by weight ratio:

-   -   25-40 parts of silybin,    -   30-50 parts of phospholipid,    -   80-120 parts of Pu'er tea extract, and    -   20-30 parts of vitamin E.

It is most preferably prepared from the following bulk drugs by weightratio:

-   -   35 parts of silybin,    -   42 parts of phospholipid,    -   100 parts of Pu'er tea extract, and    -   25 parts of vitamin E.

The phospholipid of the present: invention is a phospholipid orlecithin, which is mainly composed of phosphatidylcholine, preferablysoybean phospholipid.

The role of the phospholipid in the present invention is to promote thedissolution and absorption of pharmaceuticals, silybin is apharmaceutical with low solubility and low permeability, and thephospholipid is combined therewith to fern a phospholipid complex so asto improve solubility of the silybin, thereby improving thebioavailability of the pharmaceuticals.

Described silybin and phospholipid are both known from the prior art orcommercially available. In order to better exert the efficacy of thepresent invention, the silybin of the present invention is preferablyprepared by dissolving silymarin in 80% ethanol, filtering and washingthe precipitate with 95% ethanol for three times, collecting theprecipitate. The precipitate is dissolved in anhydrous ethanol,filtered, and the filtrate is added with a certain amount of water toseparate out the precipitate, and the precipitate is collected byfiltration, dried under reduced pressure, pulverized and mixed.

The Pu'er tea extract is commercially available, preferably a DEEPURE®Pu'er tea essence, it is also possible to be prepared according to theprior art. In order to better exert the efficacy of fee presentinvention, Pu'er tea essence or Pu'er tea extract is preferably preparedaccording to the method of patents (publication No, CN101961061A,CN101961061B, CN101961425A, CN101961425B, CN101963060A, CN101961059A,CN101961059B).

For example, said Pu'er tea essence is prepared as follows:

-   Step 1, Pu'er tea leaves are decocted with 6-12 times the volume of    water for 2-4 times. 0.5-2 hours each time; extract solution is    filtered, and filtrate is concentrated under reduced pressure and    the temperature of <70° C. to the weight of tea leaves:the volume of    concentrate=1:2-1:3;-   Step 2, the concentrate is centrifuged with a centrifuge, the    centrifugate is concentrated under reduced pressure to density of    1.1-1.25 at 45-65° C., concentrated cream is spray dried or    microwave dried to obtain the final product.

Preferably, the steps are present as follows:

-   Step 1, Pu'er tea leaves are decocted with 6-12 times the volume of    vigorously boiling water for 3 times, 0.5-2 hours each time; extract    solution is filtered, and filtrate is concentrated under reduced    pressure and the temperature of <70° C. to the weight of tea    leaves:the volume of concentrate=1:2-1:3 ;-   Step 2, the concentrate is centrifuged with a tripod pendulum type    batch centrifugal, the tripod pendulum is centrifuged with a    tubular-bowl centrifuge, and the centrifugate is concentrated under    reduced pressure to density of 1.1-1.25 at 45-65° C., concentrated    cream is spray dried or microwave dried to obtain the final product;-   wherein tubular-bowl centrifuge, condition is: centrifuge speed:    15000-19000 rpm/min; spray drying conditions are: inlet temperature:    140-190° C., outlet temperature: 75-95° C.,

Most preferably, the steps are present as follows:

Pu'er tea leaves are decocted with vigorously boiling water for 3 times,the first time decocted for 1.5 h, 10 times the volume of water added;the second time decocted for 1.5 h, 8 times the volume of water added;the third time decocted for 1 h, 8 times the volume of water added,extract solution is filtered, and filtrate is concentrated under reducedpressure and the temperature of <70° C. to the weight of tea leaves:thevolume of concentrate=1:2-1:3, the concentrate is centrifuged with atripod pendulum type batch centrifugal, the tripod pendulum iscentrifuged with a tubular-bowl centrifuge, and the centrifugate isconcentrated under reduced pressure to density of 1.1-1.25 at 45-65° C.,concentrated cream is spray dried or microwave dried to obtain the finalproduct;

-   wherein tubular-bowl centrifuge condition is: centrifuge speed:    15000-19000 rpm/min: spray drying conditions are: inlet temperature:    140-190° C., outlet temperature: 75-95° C.

The vitamin E is any one of commercially available vitamin E for dietarysupplementation or drug use, vitamin E acetate and vitamin E succinate.

The above compositions are made by weight ratios, and may be increasedor reduced according to corresponding proportion in productionprocesses, such as large-scale production can be in unit of kg or T(ton); small scale preparations can also be in unit of g. The weight canbe increased or reduced, but the proportions of the weight ratio of bulkdrugs between the components remain unchanged.

The proportions of the above weight ratio are obtained throughscientific screening, for special patients, such as patients with severeor mild symptom, obese or thin patients, the proportions of the amountof composition can be accordingly adjusted, increased or decreased nomore than 10%, the efficacy is substantially constant.

Any pharmaceutically acceptable dosage forms can be formulated in theformulation of a pharmaceutical preparation, the dosage forms areselected from: tablet, sugar coated tablet, film coated tablet, entericcoated tablet, capsule, hard capsule, soft capsule, oral liquid, oralagent, granule, pill, powder, paste, sublimed preparation, suspensionagent, solution, injection, suppository, ointment, emplastrum, creme,spray, patch. Preferably oral preparations, and optimal preferablytablet, capsule, granule.

Some pharmaceutically acceptable carriers can be added into thecompositions of the present invention as needed, the pharmaceuticalpreparations can be prepared using galenic pharmacy conventionaltechniques, such as inking the pharmaceutically active substances withpharmaceutically acceptable carriers. The pharmaceutically acceptable,carriers are selected from: mannitol, sorbitol, sorbic acid or sylvite,sodium metabisulfate, sodium bisulfite, sodium thiosulfate, cysteinehydrochloride, mercaptoaeetic acid, methionine, vitamin A, vitamin C,vitamin E, vitamin D, azone, disodium EDTA, calcium disodium EDTA, thecarbonate, acetate, phosphate of monovalence alkali metal or aqueoussolution thereof, hydrochloric acid, acetic acid, sulfuric acid,phosphoric acid, amino acid, sodium chloride, potassium chloride, sodiumlactate, xylitol, maltose, glucose, fructose, dextran, glycine, starch,sucrose, lactose, mannitol, silicon derivative, cellulose and derivatethereof, alginate, gelatin, polyvinyl pyrrolidone, glycerine, propyleneglycol, ethanol, Tween 60-80, Span-80, beeswax, lanolin, liquidparaffin, cetyl alcohol, gallic acid esters, agar, triethanolamine,basic amino acid, urea, allantom, calcium carbonate, calciumbicarbonate, surfactant, polyethylene glycol, cyclodextrin,beta-cyclodextrin, phospholipid material, kaolin, talc, calciumstearate, magnesium stearate, etc. Preferably, the carrier is one ormore of microcrystalline cellulose, lactose, starch, sodiumcarboxymethylcellulose, low substituted hydroxypropyl cellulose, talc.

When the composition of the present invention is prepared intomedicament, the unit dosage of fee medicament may contain 0.1-1,000 mgof the pharmaceutically active substance of the present invention, andthe remainders are pharmaceutically acceptable carriers. Thepharmaceutically acceptable camera may be 0.1-99.9% of the totalpreparation weight by weight, Preferably, the pharmaceuticallyacceptable earners may be 40-70% of the total preparation weight byweight.

The usage and dosage of the Chinese traditional medicine compositions orpreparations of the present invention are determined according to theconditions of patients while being used.

The cumulative dissolution rate of the pharmaceutical compositionpreparations according to the present invention, such as tablets,capsules, granules and so on, is not less than 60% while dissolution invitro for 2 h and the dissolution rate is greater than or equal to 15%while dissolution in vitro for 30 min, in a dissolution condition:slurry method, rotation speed of 100 rpm and temperature of 37 □, andrelease medium is: 1000 ml of hydrochloric acid solution at pH 1.2,dosage: 1 capsule/1 tablet/1 bag granules.

A preparation method of the pharmaceutical composition of the presentinvention comprises the following steps:

-   {circle around (1)} taking a prescription amount of raw materials    for later use;-   {circle around (2)} preparation of silybin complex liquid; weighing    a prescription amount of silybin and phospholipid, and dissolving    them in the anhydrous ethanol, heating and refluxing to clarify the    solution and continuing to heat for a certain time, then    concentrating the clear solution under reduced pressure to a certain    volume, to obtain the silybin complex liquid for later use;-   {circle around (3)} granulation: weighing a prescription amount of    Pu'er tea extract as a base material, taking the silybin complex    liquid prepared in step {circle around (2)} as a feed liquid, and    preparing granules by a fluidization spray method with a fluidized    bed, and drying after the liquid complex is all sprayed in for later    use;-   {circle around (4 )} total blending: mixing vitamin E and the    granules of step {circle around (3)} uniformly to obtain the    pharmaceutical composition;

The present invention also includes a preparation step {circle around(5)}, taking the pharmaceutical composition of step {circle around (4)}and pharmaceutically acceptable carriers, and preparing pharmaceuticallyacceptable dosage forms according to the conventional preparationprocess.

Further preferably, the preparation method of the pharmaceuticalcomposition of the present invention, comprises the following steps:

-   {circle around (1 )} taking a prescription amount of raw materials    for later use;-   {circle around (2)} preparation of silybin complex liquid: weighing    a prescription amount; of silybin and phospholipid, and dissolving    them in the anhydrous ethanol, heating and refluxing to clarify the    solution and continuing to heat for a certain time, then    concentrating the clear solution under reduced pressure to a certain    volume, to obtain the silybin complex liquid for later use;-   {circle around (3)} granulation: weighing a prescription amount of    Pu'er tea extract as a base material, taking lire silybin complex    liquid prepared in step {circle around (2 )} as a feed liquid, and    preparing granules by a fluidization spray method with a fiuidized    bed, and drying after the liquid complex is ail sprayed in for later    use;-   {circle around (4)} total blending: mixing vitamin E and the    granules of step {circle around (3)} uniformly in an equal    incremental manner to obtain the pharmaceutical composition;-   {circle around (5)} preparing pharmaceutically acceptable dosage    forms according to the conventional preparation process.

Wherein the healing time described in step {circle around (2)} is0.5-1.5 hours; the concentrated volume is 5%-20% of the original volumeand the temperature of concentration under reduced pressure is 60-80° C.

Wherein the parameters of the fluidized bed in step {circle around (3)}are: the temperature of the materials is 40-65° C., during thegranulation process, fee parameters such as fan frequency, inlet airtemperature and infusion frequency are adjusted to keep fee materials ingood fluidization state.

After completing the granulation, the granules are dried for 10-60minutes, and the drying temperature is 55-65° C.

Pu'er tea can improve insulin resistance, regulate the levels of bloodlipid and leptin and other effects, can block the fat accumulation ofhepatic parenchymal cell caused by insulin resistance to a certainextent, combined with the strong free radical scavenging andanti-oxidative stress ability of silybin, Ve is added to further enhancethe effects of the products in beautifying skin, and the three have abetter anti-NAFLD (non-alcoholic fatty liver diseases) effect as well asskin-beautifying effect.

EXPERIMENTAL EXAMPLE 1

Dissolution Experiment in vitro

The dissolutions of the silybin-phospholipid-Pu'er tea-vitamin Ecompositions obtained in embodiments 18-22 are determined under thefollowing conditions: the selection of fee dissolution methods is basedon fee properties of fee main component silybin in the compositions, thesilybin, as a medicament withe low solubility and low permeability, isthe fourth category in the Biopharmaceutics Classification System (BCS),dissolution and absorption thereof are both the rate-limiting steps,either of which should be resolved at the same time in order to improvethe bioavailability of the pharmaceuticals. Dissolution stage of thesilybin is mainly carried out in the stomach, the absorption stage ismainly carried out in the small intestine, and measuring the in-vitrodissolution of the pharmaceutical helps to improve the bioavailabilityof fee pharmaceuticals. Therefore, the following dissolution, method ischosen to evaluate the composition: slurry method, rotation speed of 100rpm and temperature of 37° C., and release medium is: 1000 ml ofhydrochloric acid solution at pH 1.2, dosage: 1 capsule/1 tablet/1 baggranules. The sampling points are: 15, 30, 45, 60, 90, 120 min. Thecumulative dissolution is determined. The results are shown in Table 1below.

TABLE 1 Accumulated dissolution (%) time Embodiment EmbodimentEmbodiment Embodiment Embodiment Embodiment Embodiment Embodiment (min)Embodiment 1 16 18 19 20 21 22 24 25 15 14.95% 12.86% 17.75% 16.45%12.86% 3.35% 7.90% 5.28% 4.98% 30 54.09% 55.65% 51.04% 54.21% 55.65%16.59% 35.66% 18.23% 14.97% 45 76.99% 67.79% 77.92% 75.98% 67.79% 49.47%62.13% 52.37% 50.09% 60 79.90% 78.43% 78.56% 79.42% 78.43% 65.13% 72.10%67.28% 66.93% 90 81.45% 80.01% 82.35% 80.13% 80.01% 78.41% 79.20% 78.09%80.28% 120 85.29% 80.86% 83.89% 80.78% 80.86% 79.69% 81.11% 82.89%81.99%

The dissolution determination of the reference preparation(silybin-phospholipid complex preparation, laboratory homemade) iscarried out, and compared with the silybin-phospholipid-Pu'ertea-vitamin E compositions prepared in embodiments 18-22, and theresults are as shown in FIG. 1. It can be seen from the data in Table 1and the graph of FIG. 1:

The in-vitro release of the silybin-phospholipid-Pu'er tea-vitamin Ecomposition prepared by the preparation method of the present inventionis significantly better than that of the reference preparationsilybin-phospholipid complex, surprisingly, the cumulative dissolutionof the composition in the hydrochloric acid solution at pH 1.2 for 2 hreaches to 80%, nearly completely dissolved, which is doubled ascompared with that of the reference preparation, resolving the problemsof low solubility and low bioavailability of silybin that have alwaysexisted, which will provide the basis for fee studies of dose settingand in-vivo safely and efficacy of silybin composition in the future.

Pooling the data, of in-vitro dissolution experiments and in-vivopharmacological researches, the present invention further improves thedissolution of the pharmaceutica by combining the silybin-phospholipidcomplex with Pu'er tea and vitamin E, by continuing the absorptionimprovements of the pharmaceutical by increasing the compatibility ofthe pharmaceutical and the biofilm after the combination of silybin andphospholipid, from two aspects of improving the dissolution andabsorption to improve the bioavailability of the main component silybin.

EXPERIMENTAL EXAMPLE 2

Evaluation of Pharmacodynamics in vivo

1 Experimental Animals

80 mice with SPF grade and 6-week-old male C57 BL/6J leptin-deficient(ob/ob), 10 mice with SPF grade and 6 weeks old male C57 BL/6J (ob/m),provided by the Beijing Huafukang Bioscience Co., Inc., raised in TaslyInstitute's pharmacological toxicology research center barrier animalroom, at the temperature of 20° C.-25° C. , relative humidity of 60%, 5mice in each cage, lighting time of 12 hours, timely and quantitativefeed, ob/ob mice are fed with high fat diet (HFD, D12492), C57 RL/6Jmice are fed with normal diet, both are provided by Beijing HuafukaogBioscience Co., Inc., and free drinking water, daily replacement ofpadding.

2 Tested Substances

Silybin-phospholipid complex, provided by Tasly Pharmaceutical Co.,Ltd., lot number 500902031; Pu'er tea essence, brown powder, provided byTasly Pharmaceutical Group Co., Ltd., lot number Z001 PE(2014)C06(H);water-soluble vitamin E, produced by Sigma Company, the above testedsubstances are stored in the sample cabinet of the test room ofPharmacology Institute to be protected from light at room temperature.

3 Experimental Methods 3.1 Experimental Dose Design and Grouping

The silybin-phospholipid complex is administered to the experimentalanimals at a daily dose of

3g (containing silybin 420 mg, soybean phospholipid 504 mg); the Pu'ertea extract is administered to the experimental animals at a daily doseof 1.2 g. The compatibility proportions and the experimental dosedesigns of the five different compositions are shown in Table 2, thedose of the experimental animals is set to the corresponding equivalentdose of the corresponding tested substances, the formula for calculationis as follows:

-   -   animal experimental dose=recommended human dose/60 kg*12.3

TABLE 2 Dose of different compositions Prescription Recommended humandaily dose (mg) pharmaceuticals Combination 1 Combination 2 Combination3 Combination 4 Combination 5 Silybin 420 420 420 105 105 phospholipid504 780 780 195 195 Pu'er tea extract 1200 1200 2400 300 1200 Vitamin E300 300 75 150 150 Prescription Experimental animal dose design (mg/kg)pharmaceuticals Combination 1 Combination 2 Combination 3 Combination 4Combination 5 Silybin 86.10 86.10 86.10 21.53 21.53 phospholipid 103.32160.00 160.00 40.00 40.00 Pu'er tea extract 246.00 246.00 492.00 61.50246.00 Vitamin E 61.50 61.50 15.38 30.75 30.75

3.2 Administration of Tested Substances

After 1 week of adaptive feeding, 80 ob/ob mice of 6-week-old arerandomly divided into 8 groups: a model group, a silybin-phospholipidcomplex group, a Pu'er tea extract group, a combination 1 group , acombination 2 group , a combination 3 group , a combination 4 group ,and a combination 5 group, 10 mice for each group. Another 10 C57BL/6 Jmice of 6-week-old are in a normal group. Normal group mice are fed withnormal diet, the model group and the administration group are fed withhigh fat diet (HFD, D12492). In addition, the mice in different drugsintervention groups are given the corresponding doses of drags by meansof intragastric administration, the doses of the five compositions areshown in Table 1, the normal group and the model group are given thesame amount of distilled wafer, continuous intragastric administrationfor 6 weeks.

The mice are free to eat and drink during the experiment, weekly weight,and the doses are adjusted according to the body weight. After the lastadministration, lasting for 12 h, cannot help but water, weighing thebody weight, extracting rats' eyeballs to collect blood and then putthem to death by breaking their necks, and the liver is harvestedrapidly, physiological saline rinsing, filter paper blotting andpreserved in a −20° C. refrigerator after weighing.

3.3 Detecting Indicators and Methods 3.3.1 General Observation

The weights of mice in each group are measured weekly during theexperiment,

3.3.2 Calculation of The Liver Index and Observation of the GeneralMorphology of the Liver

After finishing the experiment, the liver is weighed and the liver indexis calculated, the liver index (%)= liver wet weight/body weight*100%.

3.3.3Determination of Serum Biochemical Indexes

Blood of all the mice are collected by extracting rats' eyeballs andcentrifuged at 3000 r/min for 15minutes, the serum is separated andcollected in an EP tube and stored at −20° C. refrigerator for lateruse. The content of glutamic oxaloacetic transaminase (AST),glutamicpyruvic transaminase (ALT), total cholesterol (TC), low-densitylipoprotein cholesterol (LDL-C) in serum are measured by 7020 automaticbiochemistry instrument.

3.3.4Insulin Resistant Index

Serum FINS is detected using the Elisa kit and the insulin resistanceindex is calculated by the formula.

${{Home}\text{-}{IR}} = \frac{{FBG} \times {FINS}}{22.5}$

3.3.5 Liver Histopathological Examination

Frozen sections are prepared from frozen liver tissues and the degree ofhepatic steatosis is observed by oil red O staining. Oil red O stainingoperation steps; frozen slicing→sufficiently washing with distilledwater→staining with oil red O diluent in the dark tor 10-15minutes→taking out 6 ml of oil red O saturated liquid, adding 4 ml ofdistilled water, leaving it for 5-10 minutes and filtrating for lateruse→differentiating to interstitial clear under mirror with 60%ethanol→washing with water→nuclear counter staining withhematoxylin→washing with water→sealing piece with neutral gum→microscopeobservation.

3.4 Data Processing

SPSS 15.0 statistical software is used for analysis, the data areexpressed as mean±standard deviation, the t test is used to analyzewhether there's any difference between the two groups before and aftertreatment or not and the difference is statistically significant withP<0.05.

4 Experimental Results 4.1 The Effects of Each Tested Substance on BodyWeight

Mice in each group are measured once a week during the experiment, andthe effects of each group of medicaments on body weight of micesuffering from non-alcoholic fatty liver diseases in each group areobserved. As shown in Table 3, the weight of mice in normal group isincreased slowly and the weight of mice in model group is increased morerapidly. After 6 weeks of administration, the other administrationgroups except for the silybin phospholipid complex group could inhibitthe weight increases of mice in different degrees(P<0.05), a combineduse of silybin-phospholipid complex, Pu'er tea extract and vitamin E issignificantly superior to using silybin-phospholipid complex alone.

TABLE 3 Effects of each tested substance on body weight of mice (g, n =10, x ± S) Body weight (g) Before Administration AdministrationAdministration Group administration for 2 weeks for 4 weeks for 6 weeksNormal 21.35 ± 0.58 23.10 ± 1.05 24.41 ± 1.11  26.70 ± 1.56  Model 48.88± 3.27 54.98 ± 2.78 58.99 ± 3.26  61.50 ± 4.01  Silybin-phospholipidcomplex 47.19 ± 3.58 54.39 ± 4.57 58.45 ± 5.64  61.57 ± 5.61 Combination 1 46.98 ± 3.54 52.51 ± 3.12 55.80 ± 4.77  58.14 ± 3.10* Combination 2 47.50 ± 3.02  52.14 ± 2.27* 54.99 ± 2.98** 57.11 ± 3.92* Combination 3 48.25 ± 2.16  51.39 ± 2.39** 53.22 ± 2.10** 56.07 ± 3.35**Combination 4 47.67 ± 3.14  51.56 ± 1.47** 53.29 ± 2.28** 55.19 ± 3.11**Combination 5 48.52 ± 2.04  51.29 ± 2.26** 53.72 ± 2.06** 56.57 ± 3.51***Compared with the model group P < 0.05; **Compared with the model groupP < 0.01;

4.2 Effects of Each Tested Substance on Liver Index

As shown in Table 4, the body weight, liver wet weight and liver indexof mice in fee model group are significantly increased compared with feenormal group(P<0.01), and the silybin-phospholipid complex can decreasethe wet weight of fee liver (P< 0.05), but has no significant effect onbody weight and liver index of the mice (P>0.05); compatability group ofsilybin-phospholipid complex+Pu'er tea extract+VE can significantlyreduce fee wet weight and liver index of fee liver of mice (P<0.05), theeffect of a combined use of fee three is significantly better than thatof using the silybin-phospholipid complex alone.

TABLE 4 Effects of each test substance on liver index of mice Group Bodyweight (g) Liver wet weight (g) Liver index % Normal 26.20 ± 1.86  1.05± 0.13  4.02 ± 0.39  Model 61.60 ± 4.08  4.13 ± 0.43  6.70 ± 0.62 Silybin- 61.98 ± 5.43  3.79 ± 0.35*  6.15 ± 0.77  phospholipid complexCombination 1 56.37 ± 3.12** 2.74 ± 0.37** 4.86 ± 0.70*  Combination 257.39 ± 3.17*  3.50 ± 0.23** 6.01 ± 0.53*  Combination 3 57.38 ± 3.88* 2.87 ± 0.38** 5.04 ± 0.86** Combination 4 55.87 ± 2.20** 2.64 ± 0.31**4.72 ± 0.60** Combination 5 56.57 ± 3.16** 2.70 ± 0.31** 4.78 ± 0.62***Compared with the model group P < 0.05; **Compared with the model groupP < 0.01;

4.3 Effects of Each Tested Substance on Blood Lipid, Liver Function andInsulin Resistance Indexes of Mice

As shown in Table 5, the levels of serum TC, LDL, ALT, AST and insulinresistance index are significantly increased in non-alcoholic fattyliver model mice compared with the normal group (P<0.05); there is nosignificant improvement in the abnormal elevation of thesilybin-phospholipid complex (P>0.05); the silybin-phospholipid complex,Pu'er tea extract arid vitamin E compatibility group of differentproportions, except for no significant improvement in serum AST by thecombination 2 (P>0.05), the other compositions can significantly reduceTC, LDL-C, ALT, AST find insulin resistance indexes (P<0.05), and theeffect is better than that of using the silybin-phospholipid complexalone.

TABLE 5 Effects of each tested substance on blood lipid, liver functionand insulin resistance indexes of mice Insulin resistance Group TC LDL-CALT AST indexes Normal 3.10 ± 0.23  0.24 ± 0.10  58.89 ± 35.26  124.78 ±53.41  0.587 ± 0.129  Model 10.07 ± 1.08   2.11 ± 0.56  404.67 ± 102.01 190.89 ± 31.63  1.142 ± 0.227  Silybin-phospholipid complex 10.00 ±0.83   2.22 ± 0.32  431.24 ± 98.65  182.74 ± 24.86  0.988 ± 0.373 Combination 1 7.01 ± 0.99** 1.11 ± 0.21**  249.51 ± 133.14** 116.19 ±21.43** 0.689 ± 0.124** Combination 2 8.79 ± 0.68** 1.47 ± 0.22** 370.48± 76.03  149.74 ± 32.14** 0.830 ± 0.272*  Combination 3 7.95 ± 1.25**1.29 ± 0.17** 280.58 ± 141.68* 143.10 ± 18.05** 0.785 ± 0.139**Combination 4 6.34 ± 0.73** 1.02 ± 0.23** 215.47 ± 83.09** 103.97 ±10.15** 0.612 ± 0.103** Combination 5 7.12 ± 0.90** 1.15 ± 0.19** 259.51± 129.13* 118.78 ± 21.36** 0.732 ± 0.159** *Compared with the modelgroup P < 0.05; **Compared with the model group P < 0.01;

4.4 Effects of Each Tested Substance on Liver Pathology in Mice

Oil red O staining; according to the size and number of red particles inhepatocytes of liver frozen issues stained by Oil red O under lightmicroscope, it is divided into mild, moderate and severe type. Mild,that is , ⅓-⅔ of red granules are shown per unit area under lightmicroscope, graded as 1 point; moderate, that is, more than ⅔ of thehepatocytes containing red particles, graded as 2points; severe, thatis, almost all of the hepatocytes containing red particles, graded as 3points; no steatosis is observed, graded as 0 points.

As shown in Table 6, steatosis occurred in nearly all the hepatocytes inthe liver tissues of the model group, and the pathological scores aresignificantly increased than that its the normal group(P<0.01); there isno significant improvement on liver pathological scores by usingsilybin-phospholipid complex or Pu'er tea extract alone (P>0.05); thecombination of different proportions of the Pu'er tea extract,silybin-phospholipid complex and vitamin E can significantly improve theliver steatosis, reduce the pathological scores (P<0.05), and the effectis better than that of using the silybin-phospholipid complex or thePu'er tea extract alone,

TABLE 6 Effects of each tested substance on liver pathology in miceGroup Oil red O staining pathological score Normal 0.000 ± 0.000  Model2.800 ± 0.422  Silybin-phospholipid complex 2.500 ± 0.404  Pu'er teaextract 2.300 ± 0.675  Combination 1 2.000 ± 0.667** Combination 2 2.200± 0.422** Combination 3 2.200 ± 0.632*  Combination 4 1.600 ± 0.516**Combination 5 2.200 ± 0.632** *Compared with the model group P < 0.05;**Compared with the model group P < 0.01;

5 Experimental Conclusions

The above experimental results show that: the body weight, liver index,blood lipid, ALT, AST and insulin resistance index are significantlyincreased in the mice of the non-alcoholic fatty liver model groupcompared with those in the blank group, and the liver tissues are severesteatosis, Pu'er tea can improve insulin resistance, regulate bloodlipids, combined with silybin and vitamin E having strong free radicalscavenging and anti-oxidative stress ability, the combination use of thethree has a significant improvement in liver steatosis.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 is an in-vitro release curve, wherein, each sample is referencepreparation of Shui Lin Jia, Shui Lin Jia without Pu'er tea, andsilybin-phospholipid-Pu'er tea compositions prepared in embodiments16-20.

DETAILED DESCRIPTION OF THE INVENTION

The present invention is further illustrated by the following specificexamples, but not intended to limit the present invention.

Embodiment 1

Taking 26.25 g of silybin, 45 g of soybean phospholipid, 75 g of Pu'ertea extract, and 18.75 g of vitamin E.

-   {circle around (1 )} Preparation of silybin complex liquid; weighing    a prescription amount of silybin, soybean phospholipid, and    dissolving them in the anhydrous ethanol, heating and refluxing to    clarify the solution and continuing to heat for 1 h, then    concentrated under reduced pressure and recycling the ethanol to 15%    of the original volume for later use;

{circle around (2 )} Granulation: weighing a prescription amount ofPu'er tea extract as a base material, taking the silybin complex liquidprepared in step CD as a feed liquid, preparing the granules by afluidization spray method with a fiuidized bed, controlling thetemperature of materials at 40° C., drying at 60° C. for 20 min afterthe liquid complexes are all sprayed in for later use;

-   {circle around (3 )} Mixing vitamin E and the granules prepared in    step {circle around (2)} uniformly in an equal incremental manner ,    bagging, made into 1,000 bags of granules.

Embodiment 2

Taking 180 g of silybin, 195 g of soybean phospholipid, 600 g of Pu'ertea extract, and 120 g of vitamin E.

{circle around (1)} Preparation of silybin complex liquid: weighing aprescription amount of silybin, soybean phospholipid, and dissolvingthem in the anhydrous ethanol, heating and refluxing to clarify thesolution and continuing to heat for 1.5 h, then, concentrated underreduced pressure and recycling the ethanol to 20% of the original volumefor later use;

{circle around (2)} Granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking the silybin composite liquidprepared in step {circle around (1)} as a feed liquid, preparing thegranules by a fluidization spray method with a fiuidized bed,controlling the temperature of materials at 65° C., drying at 65° C. for60 min after the liquid complexes are all sprayed in for later use;

-   {circle around (3)} Mixing vitamin E and the granules prepared in    step {circle around (2 )} uniformly in an equal incremental manner,    bagging, made into 1,000 bags of granules.

Embodiment 3

Taking 26.25 g of silybin, 195 g of phospholipid, 600 g of Pu'er teaextract, and 18.75 g of vitamin E,

-   {circle around (1)} CD Preparation of silybin complex liquid:    weighing a prescription amount of silybin, soybean phospholipid, and    dissolving them in the anhydrous ethanol, heating and refluxing to    clarify the solution and continuing to heat for 0.5 h, then    concentrated under reduced pressure and recycling the ethanol to 5%    of the original volume for later use;

{circle around (2)} Granulation: weighing a prescription amount of Pu'ertea extract as a base material, taking the silybin composite liquidprepared in step {circle around (1)} as a feed liquid, preparing thegranules by a fluidization spray method with a fluidized bed,controlling the material temperature at 50° C., drying at 55° C. for 10min after the liquid complexes are ail sprayed in for later use;

-   {circle around (3)} Mixing vitamin E and the granules prepared in    step {circle around (2 )} uniformly in an equal incremental manner,    bagging, made into 1,000 bags of granules.

Embodiment 4

Taking 26.25 g of silybin, 195 g of phospholipid, 75 g of Pu'er teaextract and 120 g of vitamin E, and preparing 1,000 bags of granulesaccording to the method of Embodiment 1.

Embodiment 5

Taking 180 g of silybin, 45 g of phospholipid, 75 g of Pu'er tea extractand 18.75 g of vitamin E, and preparing 1,000 bags of granules accordingto the method of Embodiment 1.

Embodiment 6

Taking 180 g of silybin, 45 g of phospholipid, 600 g of Pu'er teaextract and 12.0 g of vitamin E, and preparing 1,000 bags of granulesaccording to the method of Embodiment 1.

Embodiment 7

Taking 180 g of silybin, 195 g of phospholipid, 75 g of Pu'er teaextract and 120 g of vitamin E, and preparing, 1,000 bags of granulesaccording to the method of Embodiment 1.

Embodiment 8

Taking 26.25 g of silybin, 48.75 g of phospholipid, 75 g of Pu'er teaextract and 37.5 g of vitamin E, and preparing 1,000 bags of granulesaccording to the method of Embodiment 1.

Embodiment 9

Taking 26.25 g of silybin, 48.75 g of phospholipid, 300 g of Pu'er teaextract and 37.5 g of vitamin E, and preparing 1,000 bags of granulesaccording to tire method of Embodiment 1.

Embodiment 10

Taking 52.5 g of silybin, 97.5 g of phospholipid, 300 g of Pu'er teaextract and 9,375 g of vitamin E, and preparing 1,000 bags of granulesaccording to the method of Embodiment 1.

Embodiment 11

Taking 75 g of silybin, 90 g of phospholipid, 240 g of Pu'er tea extractand 90 g of vitamin E, and preparing 1,000 bags of granules according tothe method of Embodiment 1.

Embodiment 12

Taking 90 g of silybin, 108 g of phospholipid, 270 g of Pu'er teaextract and 75 g of vitamin E, and preparing 1,000 bags of granulesaccording to the method of Embodiment 1.

Embodiment 13

Taking 105 g of silybin, 126 g of phospholipid, 300 g of Pu'er teaextract and 75 g of vitamin E, and preparing 1,000 bags of granulesaccording to the method of Embodiment 1.

Embodiment 14

Taking 105 g of silybin, 195 g of phospholipid, 300 g of Pu'er teaextract and 75 g of vitamin E, and preparing 1,000 bags of granulesaccording to the method of Embodiment 1.

Embodiment 15

Taking 120 g of silybin, 150 g of phospholipid, 360 g of Pu'er teaextract and 90 g of vitamin E, and preparing 1,000 bags of granulesaccording to the method of Embodiment 1.

Embodiment 16

Taking 105 g of silybin, 126 g of phospholipid, 300 g of Pu'er teaextract and 82.3 g of vitamin E acetate, and preparing 1,000 bags ofgranules according to the method of Embodiment 1.

Embodiment 17

Taking 105 g of silybin, 126 g of phospholipid, 300 g of Pu'er teaextract and 92.4 g of vitamin E succinate, and preparing 1,000 bags ofgranules according to the method of Embodiment 1.

Embodiment 18

Taking the granules of Embodiment 8, adding 549 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into No. 0 capsules to obtain 1,000 granules.

Embodiment 19

Taking the granules of Embodiment 9, adding 324 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into No. 0 capsules to obtain 1,000 granules.

Embodiment 20

Taking the granules of Embodiment 10, adding 308 g of microcrystallinecellulose and 32 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 21

Taking the granules of Embodiment 13, adding 194 g of microcrystallinecellulose, mixing uniformly, encapsulated into capsules to obtain 1,000capsules.

Embodiment 22

Taking the granules of Embodiment 14, adding 61 g of microcrystallinecellulose and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into capsules to obtain 1,000 capsules.

Embodiment 23

Taking the composition of Embodiment 8, adding 400 g of lactose, 149 gof starch and 64 g of sodium carboxymethyl starch, mixing uniformly,encapsulated into No. 0 capsules to obtain 1,000granules.

Embodiment 24

Taking the composition of Embodiment 13, adding 90 g of lactose, 10 g oftalc and 94 g of low-substituted hydroxypropyl cellulose, mixinguniformly, encapsulated into No. 0 capsules to obtain 1,000 granules.

Embodiment 25

Taking the granules of Embodiment 13, adding 194 g of microcrystallinecellulose, mixing uniformly, tablet pressing to obtain 1,000 tablets.

1. A pharmaceutical composition comprising: 8.75-60 parts by weight ofsilybin; 15-65 parts by weight of phospholipid; 25-200 parts by weightof Pu'er tea extract; and 6.25-40 parts by weight of vitamin E.
 2. Thepharmaceutical composition according to claim 1, comprising: 25-40 byweight parts of silybin; 30-50 parts by weight of phospholipid; 80-120parts by weight of Pu'er tea extract; and 20-30 parts by weight ofvitamin E.
 3. The pharmaceutical composition according to claim 1,comprising: 35 parts by weight of silybin; 42 parts by weight ofphospholipid; 100 parts by weight of Pu'er tea extract; and 25 parts byweight of vitamin E.
 4. The pharmaceutical composition according toclaim 1 wherein the vitamin E is vitamin E acetate or vitamin Esuccinate.
 5. A pharmaceutical preparation comprising the pharmaceuticalcomposition according to claim 1, and pharmaceutically acceptablecarriers; wherein the pharmaceutically acceptable carriers are 0.1-99.9%of the total preparation by weight; or wherein the pharmaceuticallyacceptable carriers may be are 40-70% of the total preparation byweight.
 6. The pharmaceutical preparation according to claim 5, whereinthe pharmaceutically acceptable carriers comprise none or one or more ofmannitol, sorbitol, sorbic acid or sylvite, sodium metabisulfite, sodiumbisulfite, sodium thiosulfate, cysteine hydrochloride, mercaptoaceticacid, methionine, vitamin A, vitamin C, vitamin E, vitamin D, azone,disodium EDTA, calcium disodium EDTA, the carbonate, acetate, phosphateof monovalence alkali metal or aqueous solution thereof, hydrochloricacid, acetic acid, sulfuric acid, phosphoric acid, amino acid, sodiumchloride, potassium chloride, sodium lactate, xylitol, maltose, glucose,fructose, dextran, glycine, starch, sucrose, lactose, mannitol, siliconderivative, cellulose and derivate thereof, alginate, gelatin, polyvinylpyrrolidone, glycerine, propylene glycol, ethanol, Tween 60-80, Span-80,beeswax, lanolin, liquid paraffin, cetyl alcohol, gallic acid esters,agar, triethanolamine, basic amino acid, urea, allantoin, calciumcarbonate, calcium bicarbonate, surfactant, polyethylene glycol,cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talc,calcium stearate, magnesium stearate, or microcrystalline; preferably,wherein the carriers comprise one or more of microcrystalline cellulose,lactose, starch, sodium carboxymethylcellulose, low-substitutedhydroxypropyl cellulose, or talc.
 7. The pharmaceutical preparationaccording to claim 5, wherein the pharmaceutical preparation is any oneof tablet, sugar coated tablet, film coated tablet, enteric coatedtablet, capsule, hard capsule, soft capsule, oral liquid, oral agent,granule, pill, powder, paste, sublimed preparation, supensoid agent,solution, injection, suppository, ointment, emplastrum, creme, spray, orpatch; preferably, wherein the pharmaceutical preparation is a capsule,granule or tablet.
 8. A method of preparing the pharmaceuticalpreparation according to claim 5, comprising: (1) taking a prescriptionamount of raw materials for later use; (2) preparing a silybin complexliquid by weighing a prescription amount of silybin and a phospholipid,and dissolving them in anhydrous ethanol, heating and refluxing toclarify the solution and continuing to heat for a time, thenconcentrating the clear solution under reduced pressure to aconcentrated volume, to obtain the silybin complex liquid for later use;(3) granulating by weighing a prescription amount of Pu'er tea extractas a base material, taking the silybin complex liquid prepared in step(2) as a feed liquid, and preparing granules by a fluidization spraymethod with a fluidized bed, and drying after the liquid complex is allsprayed in for later use; (4) achieving total blending by mixing theprescription amount of VE and the granules of step 3 uniformly to obtainthe pharmaceutical composition; (5) combining the pharmaceuticalcomposition and the pharmaceutically acceptable carriers to make aconventional preparation.
 9. The method according to claim 8, whereinthe heating time in step (2) is 0.5-1.5 hours; the concentrated volumeis 5%-20% of the original volume and the temperature of theconcentration under reduced pressure is 60-80° C.; the parameters of thefluidized bed in step 3 are that the temperature of the materials is40-65° C., and parameters such as fan frequency, the inlet airtemperature and the infusion frequency are adjusted to keep thematerials in a good fluidization state during the granulation process;and wherein after the granulation is completed, the granules are driedfor 10-60 minutes, and the drying temperature is 55-65° C.
 10. Use ofthe pharmaceutical composition according to claim 1 in the preparationof drugs for treating non-alcoholic fatty liver diseases and/or reducingfat and losing weight and beautifying skin.
 11. The pharmaceuticalcomposition according to claim 2 wherein the vitamin E is vitamin Eacetate or vitamin E succinate.
 12. The pharmaceutical compositionaccording to claim 3 wherein the vitamin E is vitamin E acetate orvitamin E succinate.
 13. A pharmaceutical preparation comprising thepharmaceutical composition according to claim 2, and pharmaceuticallyacceptable carriers; wherein the pharmaceutically acceptable carriersare 0.1-99.9% of the total preparation by weight; or wherein thepharmaceutically acceptable carriers are 40-70% of the total preparationby weight.
 14. The pharmaceutical preparation according to claim 13,wherein the pharmaceutically acceptable carriers comprise one or more ofmannitol, sorbitol, sorbic acid or sylvite, sodium metabisulfite, sodiumbisulfite, sodium thiosulfate, cysteine hydrochloride, mercaptoaceticacid, methionine, vitamin A, vitamin C, vitamin E, vitamin D, azone,disodium EDTA, calcium disodium EDTA, the carbonate, acetate, phosphateof monovalence alkali metal or aqueous solution thereof, hydrochloricacid, acetic acid, sulfuric acid, phosphoric acid, amino acid, sodiumchloride, potassium chloride, sodium lactate, xylitol, maltose, glucose,fructose, dextran, glycine, starch, sucrose, lactose, mannitol, siliconderivative, cellulose and derivate thereof, alginate, gelatin, polyvinylpyrrolidone, glycerine, propylene glycol, ethanol, Tween 60-80, Span-80,beeswax, lanolin, liquid paraffin, cetyl alcohol, gallic acid esters,agar, triethanolamine, basic amino acid, urea, allantoin, calciumcarbonate, calcium bicarbonate, surfactant, polyethylene glycol,cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talc,calcium stearate, magnesium stearate, or microcrystalline; preferably,wherein the carriers comprise one or more of microcrystalline cellulose,lactose, starch, sodium carboxymethylcellulose, low-substitutedhydroxypropyl cellulose, or talc.
 15. The pharmaceutical preparationaccording to claim 13, wherein the pharmaceutical preparation is any oneof tablet, sugar coated tablet, film coated tablet, enteric coatedtablet, capsule, hard capsule, soft capsule, oral liquid, oral agent,granule, pill, powder, paste, sublimed preparation, supensoid agent,solution, injection, suppository, ointment, emplastrum, creme, spray, orpatch; preferably, wherein the pharmaceutical preparation is a capsule,granule or tablet.
 16. A pharmaceutical preparation comprising thepharmaceutical composition according to claim 3, and pharmaceuticallyacceptable carriers; wherein the pharmaceutically acceptable carriersare 0.1-99.9% of the total preparation by weight; or wherein thepharmaceutically acceptable carriers are 40-70% of the total preparationby weight.
 17. The pharmaceutical preparation according to claim 16,wherein the pharmaceutically acceptable carriers comprise one or more ofmannitol, sorbitol, sorbic acid or sylvite, sodium metabisulfite, sodiumbisulfite, sodium thiosulfate, cysteine hydrochloride, mercaptoaceticacid, methionine, vitamin A, vitamin C, vitamin E, vitamin D, azone,disodium EDTA, calcium disodium EDTA, the carbonate, acetate, phosphateof monovalence alkali metal or aqueous solution thereof, hydrochloricacid, acetic acid, sulfuric acid, phosphoric acid, amino acid, sodiumchloride, potassium chloride, sodium lactate, xylitol, maltose, glucose,fructose, dextran, glycine, starch, sucrose, lactose, mannitol, siliconderivative, cellulose and derivate thereof, alginate, gelatin, polyvinylpyrrolidone, glycerine, propylene glycol, ethanol, Tween 60-80, Span-80,beeswax, lanolin, liquid paraffin, cetyl alcohol, gallic acid esters,agar, triethanolamine, basic amino acid, urea, allantoin, calciumcarbonate, calcium bicarbonate, surfactant, polyethylene glycol,cyclodextrin, beta-cyclodextrin, phospholipid material, kaolin, talc,calcium stearate, magnesium stearate, or microcrystalline; preferably,wherein the carriers comprise one or more of microcrystalline cellulose,lactose, starch, sodium carboxymethylcellulose, low-substitutedhydroxypropyl cellulose, or talc.
 18. A method of preparing thepharmaceutical preparation according to claim 13, comprising: (1) takinga prescription amount of raw materials for later use; (2) preparing asilybin complex liquid by weighing a prescription amount of silybin anda phospholipid, and dissolving them in anhydrous ethanol, heating andrefluxing to clarify the solution and continuing to heat for a time,then concentrating the clear solution under reduced pressure to aconcentrated volume, to obtain the silybin complex liquid for later use;(3) granulating by weighing a prescription amount of Pu'er tea extractas a base material, taking the silybin complex liquid prepared in step(2) as a feed liquid, and preparing granules by a fluidization spraymethod with a fluidized bed, and drying after the liquid complex is allsprayed in for later use; (4) achieving total blending by mixing theprescription amount of VE and the granules of step 3uniformly to obtainthe pharmaceutical composition; (5) combining the pharmaceuticalcomposition and the pharmaceutically acceptable carriers to make aconventional preparation.
 19. The method according to claim 18, whereinthe heating time in step (2) is 0.5-1.5 hours; the concentrated volumeis 5%-20% of the original volume and the temperature of theconcentration under reduced pressure is 60-80° C.; the parameters of thefluidized bed in step 3 are that the temperature of the materials is40-65° C., and parameters such as fan frequency, the inlet airtemperature and the infusion frequency are adjusted to keep thematerials in a good fluidization state during the granulation process;and wherein after the granulation is completed, the granules are driedfor 10-60 minutes, and the drying temperature is 55-65° C.
 20. Use ofthe pharmaceutical preparation according to claim 5 in the preparationof drugs for treating non-alcoholic fatty liver diseases and/or reducingfat and losing weight and beautifying skin.